孔雀草茎、叶化学成分的分离鉴定

目的:孔雀草Tagetes patula是菊科Asteraceae植物,近代研究发现花中含有大量叶黄素具有抗氧化活性,根中得到的噻吩类衍生物具有抗菌活性,根部甲醇提取物中分得的柠檬酸、苹果酸具有降压活性,该文旨在研究其茎、叶化学成分,揭示其治疗疾病的物质基础。方法:采用95%乙醇,减压回收溶剂得浓缩物,将浓缩物溶于水中,用乙酸乙酯萃取。取乙酸乙酯萃取物,用硅胶柱、开放ODS柱及制备HPLC等色谱技术进行分离纯化,根据化合物的理化性质,1H-NMR,13C-NMR等光谱数据进行结构鉴定。结果:从乙酸乙酯萃取部分中分离得到8个化合物,经光谱分析确定其结构分别为丁香脂素-4'-O-β-D-葡萄糖苷(1),2-甲氧基-4-(2-丙烯基)苯基-β-D-葡萄糖苷(2),邻苯二甲酸二丁酯(3),万寿菊素(4),β-胡萝卜苷(5),β-谷甾醇(6),4-烯丙基-2,6-二甲氧基苯基葡萄糖苷(7),1-β-D-吡喃葡萄糖苷-2,6-二甲氧基-4-丙烯基苯酚(8)。结论:化合物1,2,3,7,8为首次从孔雀草中分离得到,同时也是首次从万寿菊属植物中分离得到。该研究为综合开发及寻找天然植物性抗炎、抗氧化活性成分提供一定的化学依据和基础。     

万寿菊素和槲皮万寿菊素的分离和结构鉴定

利用有机溶剂提取及重结晶法,从菊科万寿菊属植物孔雀草(Tagetes patula.L.)分离出黄酮类化合物结晶Ⅰ;从万寿菊(Tagetes erectaL.)分离出黄酮类化合物晶Ⅱ。经化学和光谱分析,证实晶Ⅰ是万寿菊素 patuletin、晶Ⅱ是槲皮万寿菊素 quercetagetin。药理试验表明,这两种黄酮类化合物均对大白鼠晶体醛糖还原酶(AR)具有一定抑制作用。     

黑龙江产万寿菊花醇提物的指纹图谱研究

目的建立黑龙江产万寿菊花醇提物的指纹图谱并定性分析共有峰,为万寿菊花的质量控制提供参考。方法采用HPLC分离UV检测建立色谱指纹图谱,Zorbax Eclipse XDB C18色谱柱(250 mm×4.6 mm,5μm),乙腈-0.2%磷酸水溶液梯度洗脱,体积流量1 mL/min,柱温25℃,检测波长254 nm;用LC-MS定性共有峰,电喷雾离子源(ESI),负离子模式扫描,雾化器压力241.325 kPa,干燥器温度350℃,干燥气体积流量10 L/min,扫描范围:m/z 50～1 000。结果万寿菊花醇提物的指纹图谱有8个共有峰,并具有较好的精密度、重复性和稳定性,其中5个共有峰所对应的化学物质为万寿菊素、异槲皮素、异槲皮素苷和槲皮万寿菊素及其苷。结论建立的万寿菊花醇提物的指纹图谱,为万寿菊花的质量控制提供参考。     

Bioassay-guided isolation of antioxidant agents with analgesic properties from flowers of Tagetes patula

Context: Tagetes patula L. is one of the French marigold group of the Asteraceae family. It is recognized in folklore for its medicinal and pesticidal properties.

Objective: In search of more effective, but non-toxic compounds with antioxidative potential led to the bioassay guided isolation studies on the extracts of T. patula.

Materials and methods: The bioassay on Tagetes patula flowers were carried out guided by in vitro antioxidant activity using DPPH assay. A minor but proven plant constituent methyl protocatechuate (1) was isolated by column chromatography, while patuletin (2) and patulitrin (3) obtained in bulk by employing solvent partition of methanol extract. Derivatization of patuletin into benzoyl, cinnamoyl and methyl was conducted to establish the structure activity relationship (SAR). Analgesic activity of compound 2 was evaluated using acetic acid-induced writhing test and hot-plate test in mice. The toxicity of methanol extract and compound 2 were also determined.

Results: Polar extracts, fractions and phases demonstrated better antioxidant activity. The synthetic methyl protocatechuate (1) showed IC₅₀ value of 2.8?±?0.2?μg/mL, whereas patuletin (2) (IC₅₀?=?4.3?±?0.25 µg/mL) was comparable to quercetin and rutin but significantly better than patulitrin (3) (IC₅₀?=?10.17?±?1.16 µg/mL). Toxicity test for the methanol extract and compound 2 did not elicit any behavioral changes or cause mortality in mice. Compound 2 also demonstrated mild analgesic property.

Discussion and conclusion: These findings demonstrate that the plant polar extracts and fractions possess significant antioxidant property with non-toxic effect. Compound 1 is a genuine plant constituent of T. patula.     

NMR and Radical Scavenging Activities of Patuletin from Urtica urens. Against Aflatoxin B1

Abstract

In a previous study, we reported six flavonoids isolated from the methanol extract of Urtica urens L. Patuletin, the major component, was found to have powerful anti-inflammatory and antimicrobial activity. We have now investigated whether patuletin also has antioxidant activity and free radical scavenging effects in rats treated with aflatoxin B₁ (AFB₁). Male Sprague-Dawley rats (60) were assigned to 6 experimental groups including a control group and groups treated for 10 days with patuletin at low dose (7.5 mg/kg b.w) or high dose (10 mg/kg b.w) with or without AFB₁ (2 mg/kg b.w). Blood samples were collected at the end of the experimental period for biochemical analysis. The results showed significant changes typical to aflatoxicosis in rats treated only with AFB₁. Patuletin at the two tested doses caused an increase in glutathione peroxidase (GPx) and superoxide dismutase (SOD); high doses also caused an increase in liver and kidney enzymes, except total bilirubin (TB), which was in the normal value of the control animals. On the other hand, these enzymes were comparable to the controls in rats treated with low doses. Cotreatment of AFB₁ and patuletin resulted in a significant improvement in all parameters tested. We conclude that patuletin possesses an antioxidant activity and free radical scavenging effects in AFB₁-treated rats that can be observed at a dose as low as 7.5 mg/kg b.w.     

Antibacterial and Antifungal Activities of Different Parts of Tagetes patula.: Preparation of Patuletin Derivatives

Abstract

The current study evaluates the antibacterial and antifungal activities of extracts from different parts of Tagetes patula. Linn. (Asteraceae), reported for the first time in a single set of experiments. In the preliminary assay, the methanol extract of the flower (JFM) was found to possess antimicrobial activity against a number of bacteria with inhibition zone diameters ranging from 9 to 20 mm, the bioassay-guided fractionation of which led to the isolation of a flavonoid patuletin (3) in high yield as the active antibacterial principle with minimum inhibitory concentration (MIC) value of 12.5 μ g/disk against Corynebacterium. spp., Staphylococcus. spp., Streptococcus. spp., and Micrococcus luteus.. Its glucoside, patulitrin (4), was found to be weakly active, except against Staphylococcus saprophyticus., Streptococcus fecalis., and Streptococcus pyogenes. with inhibition zone diameters of 11, 16, and 12 mm, respectively. The cinnamate derivative (3b) of 3 showed antibacterial activity comparable with the parent flavonoid with a MIC value of 50 μ g/disk against Corynebacterium. spp., whereas benzoate derivative (3a) was found to be devoid of any activity; both the derivatives are new compounds. Moreover, the long-chain alcohol 5, which displayed antibacterial activity in the preliminary testing, was obtained in large quantity directly from the petroleum ether extract of the involucre of the flowers.     

黑龙江产万寿菊花醇提物的指纹图谱研究

目的 建立黑龙江产万寿菊花醇提物的指纹图谱并定性分析共有峰,为万寿菊花的质量控制提供参考。方法 采用HPLC分离UV检测建立色谱指纹图谱,Zorbax Eclipse XDB C₁₈色谱柱（250 mm×4.6 mm,5 μm）,乙腈-0.2%磷酸水溶液梯度洗脱,体积流量1 mL/min,柱温25 ℃,检测波长254 nm;用LC-MS定性共有峰,电喷雾离子源（ESI）,负离子模式扫描,雾化器压力241.325 kPa,干燥器温度350 ℃,干燥气体积流量10 L/min,扫描范围：m/z 50～1 000。结果 万寿菊花醇提物的指纹图谱有8个共有峰,并具有较好的精密度、重复性和稳定性,其中5个共有峰所对应的化学物质为万寿菊素、异槲皮素、异槲皮素苷和槲皮万寿菊素及其苷。结论 建立的万寿菊花醇提物的指纹图谱,为万寿菊花的质量控制提供参考。